| Micro Neutralization (MN) Assay of Influenza Viruses with Monoclonal Antibodies
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Author:
Ying Wu, MyungSam Cho, David Shore, Manki Song, JungAh Choi, Tao Jiang, Yong-Qiang Deng, Melissa Bourgeois, Lynn Almli, Hua Yang, Li-Mei Chen, Yi Shi, Jianxu Qi, An Li, Kye Sook Yi, MinSeok Chang, Jin Soo Bae, HyunJoo Lee, JiYoung Shin, James Stevens, SeoungSuh Hong, Cheng-Feng Qin, George F. Gao, Shin Jae Chang and Ruben O. Donis,
Date:
2016-06-05
[Abstract] The human monoclonal antibodies generated from single human B cells were tested to characterize their ability to neutralize virus infectivity. The microneutralization assay is a highly sensitive and specific assay for detecting virus-specific neutralizing antibodies to influenza viruses. This protocol is to measure the ability of human monoclonal antibody to neutralize influenza virus by microneutralization assay.
[摘要] 测试从单个人B细胞产生的人单克隆抗体，以表征其中和病毒感染性的能力。 微量中和测定法是用于检测流感病毒的病毒特异性中和抗体的高度灵敏和特异性的测定法。 该方案是测量人单克隆抗体通过微量中和测定中和流感病毒的能力。
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| Helicase Assays
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Author:
Date:
2014-03-20
[Abstract] Helicases are a class of enzymes which are motor proteins using energy derived from ATP hydrolysis to move directionally along a nucliec acid phosphodiester backbone (such as DNA, RNA and DNA-RNA hybrids) and separate two annealed nucleic acid strands. Many cellular processes, such as transcription, DNA replication, recombination and DNA repair involve helicase activity. Here, we provide a protocol to analyze helicase activities in vitro. In this protocol, the DNA helicase protein Merkel cell polyomavirus large T-antigen was expressed in the mammalian cell line HEK293 and immoblized on an IgG resin. The helicase assay is performing while the protein is immoblized on IgG resin.
[摘要] 解旋酶是一类酶，其是使用来自ATP水解的能量沿着核酸磷酸二酯主链（例如DNA，RNA和DNA-RNA杂交体）定向移动并分离两条退火的核酸链的运动蛋白。 许多细胞过程，例如转录，DNA复制，重组和DNA修复涉及解旋酶活性。 在这里，我们提供了一个协议，以分析解旋酶活性在体外。 在该协议中，DNA解旋酶蛋白Merkel细胞多瘤病毒大T抗原在哺乳动物细胞系HEK293中表达并固定在IgG树脂上。 进行解旋酶测定，同时蛋白质固定在IgG树脂上。
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| HIV-1 Virus-like Particle Budding Assay
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Author:
Date:
2013-07-20
[Abstract] Viral replication culminates with the egress of the mature virion from the host cell. This step of the viral life cycle has recently garnered increased attention with the discovery of the cellular restriction factor, Tetherin, which tethers budded virions to the surface of infected cells and inhibits viral spread. The importance of this block in viral infections has been suggested by the discovery of viral antagonists, such as HIV-1 Vpu, which counteract Tetherin. This protocol describes a system to study HIV-1 budding under BSL-2 safety conditions. It takes advantage of the ability of many viral matrix/capsid proteins to generate non-infectious virus-like particles (VLPs) with the expression of a single viral protein (i.e. HIV-1 p24 Gag). This protocol was recently used to ...
[摘要] 病毒复制随着来自宿主细胞的成熟病毒体的流出而达到高潮。病毒生命周期的这个步骤最近通过发现细胞限制因子Tetherin而发现增加的注意力，Tetherin将暴露的病毒粒子束缚于感染细胞的表面并抑制病毒扩散。这种阻断在病毒感染中的重要性已经通过发现病毒拮抗剂如HIV-1 Vpu（其抵抗了细胞因子）而提出。该协议描述了在BSL-2安全条件下研究HIV-1芽生的系统。它利用了许多病毒基质/衣壳蛋白产生具有单个病毒蛋白（即HIV-1p24Gag）表达的非感染性病毒样颗粒（VLPs）的能力。这个协议最近用于表征在HIV-1 Vpu存在下，Tetherin亚型对VLP释放的影响（Cocka和Bates，2012）。同时表达的Tetherin和其他病毒拮抗剂可用于研究Tetherin介导的病毒芽出的限制。
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